High-purity texaphyrin metal complexes

ABSTRACT

Disclosed herein are the methods and compositions for the improved synthesis of texaphyrin metal complexes. The improved synthesis results in high-purity compositions of texaphyrin metal complexes in which more than about 98% of the texaphyrin metal complexes in the composition have the same structure and/or the same molecular weight. Further described herein are pharmaceutical compositions comprising such high-purity compositions, and the use of such high-purity compositions in the treatment of cancer and cardiovascular diseases and disorders.

FIELD OF THE INVENTION

Disclosed herein are high-purity texaphyrin metal complexes, methods formaking such complexes, and the use of such high-purity texaphyrin metalcomplexes to treat diseases, disorders and conditions, including cancerand atherosclerosis.

BACKGROUND OF THE INVENTION

Cancer is a serious threat to modern society. Worldwide, more than 10million people are diagnosed with cancer every year and it is estimatedthat this number will grow to 15 million new cases every year by 2020.Cancer causes six million deaths every year or 12% of the deathsworldwide. Current treatment options are often limited but widelyemployed. Of the 1.2 million patients newly diagnosed with cancer in theUnited States annually, approximately 50% will be treated with radiationtherapy as part of initial disease management. Approximately 150,000additional patients with recurrent cancer may receive radiation therapyeach year in the U.S. Chemotherapy is administered to about 350,000cancer patients in the U.S. annually.

High levels of circulating cholesterol are associated withatherosclerosis, which may result in life-threatening blockages of bloodvessels to the heart and brain. Unstable angina, myocardial infarction(heart attack) and sudden ischemic death remain the leading cause ofmorbidity and mortality in developed nations. Current estimates indicatethat 1.1 million people in the U.S. annually will have a new orrecurrent coronary attack, and over 45 percent of these patients may diefrom the coronary attack.

Texaphyrins are small molecules having a ring-shaped chemical structurecontaining one of several metal atoms. The physical and chemicalcharacteristics of texaphyrin molecules are determined by the propertiesof the ring and the type of metal atom inserted into the ring.Texaphyrins are designed to selectively concentrate in diseased tissuesuch as tumor cells and atherosclerotic plaque inside blood vessels.Inside diseased cells, texaphyrins block crucial steps in cellularmetabolism and disrupt bioenergetic processes. Texaphyrins are designedto provide a valuable therapeutic approach to a broad range of diseases.These can be used for the treatment of a variety of diseases, includingcancer, atherosclerosis and cardiovascular diseases, and potentiallyneurodegenerative diseases, inflammatory diseases, and HIV/AIDS.

SUMMARY OF THE INVENTION

Disclosed herein are compositions comprising high-purity texaphyrinmetal complexes, wherein the identity of the substituents on thetexaphyrin complex are at least about 98% identical on all texaphyrincomplexes in a sample. Also disclosed herein are compositions comprisinghigh-purity synthetic precursors to high-purity texaphyrin complexes.Also disclosed herein are methods for synthesizing such high-puritytexaphyrin metal complexes and such high-purity precursors. Alsodisclosed herein are methods for pegylating an aromatic moiety, whereinthe identity of the poly(ethylene glycol) substituents on the aromaticmoiety are at least about 98% identical on all aromatic moieties in asample. Also disclosed herein are pharmaceutical compositions comprisingsuch high-purity texaphyrin metal complexes. Also disclosed herein aremethods for treating a disorder, disease or condition in a human patientcomprising administering to the patient an effective amount of suchhigh-purity texaphyrin metal complexes.

A first aspect disclosed herein are high-purity compositions comprisinga compound of Formula 1

wherein: M is a trivalent metal cation selected from the groupconsisting of Gd⁺³, and Lu⁺³; each X is independently selected from thegroup consisting of OH⁻, AcO⁻, Cl⁻, Br⁻, I⁻, F⁻, H₂PO₄ ⁻, ClO⁻, ClO₂ ⁻,ClO₃ ⁻, ClO₄ ⁻, HCO₃ ⁻, HSO₄ ⁻, NO₃ ⁻, N₃ ⁻, CN⁻, SCN⁻, and OCN⁻; R₃,R₄, R₅, R₆, R₇ and R₈ are independently H, OH, C_(n)H_((2n+1))O_(y) orOC_(n)H_((2n+1))O_(y) and R₁, R₂ are independently H or C₁-C₆ alkylwhere at least one of R₃, R₄, R₅, R₆, R₇ and R₈ is C_(n)H_((2n+1))O_(y)or OC_(n)H_((2n+1))O_(y), having at least one hydroxyl substituent; n isa positive integer from 1 to 11; y is zero or a positive integer lessthan or equal to n; each x is independently selected from the groupconsisting of 2, 3, 4, 5, and 6; wherein at least about 98.4% ofcompounds of Formula 1 in the composition have the same structure. Inone embodiment, M is Gd⁺³. In one embodiment, R₄ and R₇ are C₃H₆OH; R₅and R₆ are C₂H₅; R₃ and R₈ are CH₃; R₁ and R₂ are H. In one embodiment,each x is 3. In one embodiment, each X is AcO⁻. In another embodiment, Mis Lu⁺³. In one embodiment, R₄ and R₇ are C₃H₆OH; R₅ and R₆ are C₂H₅; R₃and R₈ are CH₃; R₁ and R₂ are H. In one embodiment, each x is 3. In oneembodiment, each X is AcO⁻. In a further or alternate embodiment, each Xis selected from the group consisting of sugar derivatives, cholesterolderivatives, PEG acids, organic acids, organosulfates, organophosphates,phosphates or inorganic ligands. In a further or alternate embodiment, Xis derived from an acid selected from the group consisting of gluconicacid, glucoronic acid, cholic acid, deoxycholic acid, methylphosphonicacid, phenylphosphonic acid, phosphoric acid, formic acid, propionicacid, butyric acid, pentanoic acid, 3,6,9-trioxodecanoic acid,3,6-dioxoheptanoic acid, 2,5-dioxoheptanoic acid, methylvaleric acid,glycolic acid, pyruvic acid, oxalic acid, malic acid, malonic acid,succinic acid, maleic acid, fumaric acid, tartaric acid, citric acid,methanesulfonic acid, ethanesulfonic acid, benzoic acid, salicylic acid,3-fluorobenzoic acid, 4-aminobenzoic acid, cinnamic acid, mandelic acid,and p-toluene-sulfonic acid.

In one embodiment, at least about 98.4% of compounds of Formula 1 in thecomposition have the same molecular weight, wherein the same molecularweight excludes isotopic variations. In one embodiment, at least about98.7% of compounds of Formula 1 in the composition have the samemolecular weight, wherein the same molecular weight excludes isotopicvariations. In one embodiment, at least about 99% of compounds ofFormula 1 in the composition have the same molecular weight, wherein thesame molecular weight excludes isotopic variations. In one embodiment,at least about 99.3% of compounds of Formula 1 in the composition havethe same molecular weight, wherein the same molecular weight excludesisotopic variations. In one embodiment, at least about 99.5% ofcompounds of Formula 1 in the composition have the same molecularweight, wherein the same molecular weight excludes isotopic variations.In one embodiment, at least about 98.7% of compounds of Formula 1 in thecomposition have the same structure. In one embodiment, at least about99% of compounds of Formula 1 in the composition have the samestructure. In one embodiment, at least about 99.3% of compounds ofFormula 1 in the composition have the same structure. In one embodiment,at least about 99.5% of compounds of Formula 1 in the composition havethe same structure.

A second aspect disclosed herein are high-purity compositions comprisinga compound of Formula 2:

wherein: R₃, R₄, R₅, R₆, R₇ and R₈ are independently H, OH,C_(n)H_((2n+1))O_(y) or OC_(n)H_((2n+1))O_(y) and R₁, R₂ areindependently H or C₁-C₆ alkyl where at least one of R₃, R₄, R₅, R₆, R₇and R₈ is C_(n)H_((2n+1))O_(y) or OC_(n)H_((2n+1))O_(y), having at leastone hydroxyl substituent; n is a positive integer from 1 to 11; y iszero or a positive integer less than or equal to n; each x isindependently selected from the group consisting of 2, 3, 4, 5, and 6;and acceptable acid addition salts thereof; wherein at least about 98.4%of compounds of Formula 2 in the composition have the same structure. Inone embodiment, R₄ and R₇ are C₃H₆OH; R₅ and R₆ are C₂H₅; R₃ and R₈ areCH₃; R₁ and R₂ are H. In one embodiment, each x is 3. In one embodiment,the acceptable acid addition salt is selected from the group consistingof hydrochloric acid, hydrobromic acid, sulfuric acid, nitric acid,phosphoric acid, methanesulfonic acid, ethanesulfonic acid,p-toluenesulfonic acid, and salicylic acid. In one embodiment, theacceptable acid addition salt is a hydrochloride salt.

In one embodiment, at least about 98.4% of compounds of Formula 2 in thecomposition have the same molecular weight, wherein the same molecularweight excludes isotopic variations. In one embodiment, at least about98.7% of compounds of Formula 2 in the composition have the samemolecular weight, wherein the same molecular weight excludes isotopicvariations. In one embodiment, at least about 99% of compounds ofFormula 2 in the composition have the same molecular weight, wherein thesame molecular weight excludes isotopic variations. In one embodiment,at least about 99.3% of compounds of Formula 2 in the composition havethe same molecular weight, wherein the same molecular weight excludesisotopic variations. In one embodiment, at least about 99.5% ofcompounds of Formula 2 in the composition have the same molecularweight, wherein the same molecular weight excludes isotopic variations.In one embodiment, at least about 98.7% of compounds of Formula 2 in thecomposition have the same structure. In one embodiment, at least about99% of compounds of Formula 2 in the composition have the samestructure. In one embodiment, at least about 99.3% of compounds ofFormula 2 in the composition have the same structure. In one embodiment,at least about 99.5% of compounds of Formula 2 in the composition havethe same structure.

A third aspect disclosed herein are high-purity compositions comprisinga compound of Formula 4

wherein R₁ and R₂ are independently H or C₁-C₆ alkyl; each x isindependently selected from the group consisting of 2, 3, 4, 5, and 6;and wherein at least about 98.4% of compounds of Formula 4 in thecomposition have the same structure. In one embodiment, R₁ and R₂ are H.In one embodiment, each x is 3.

In one embodiment, at least about 98.4% of compounds of Formula 4 in thecomposition have the same molecular weight, wherein the same molecularweight excludes isotopic variations. In one embodiment, at least about98.7% of compounds of Formula 4 in the composition have the samemolecular weight, wherein the same molecular weight excludes isotopicvariations. In one embodiment, at least about 99% of compounds ofFormula 4 in the composition have the same molecular weight, wherein thesame molecular weight excludes isotopic variations. In one embodiment,at least about 99.3% of compounds of Formula 4 in the composition havethe same molecular weight, wherein the same molecular weight excludesisotopic variations. In one embodiment, at least about 99.5% ofcompounds of Formula 4 in the composition have the same molecularweight, wherein the same molecular weight excludes isotopic variations.In one embodiment, at least about 98.7% of compounds of Formula 4 in thecomposition have the same structure. In one embodiment, at least about99% of compounds of Formula 4 in the composition have the samestructure. In one embodiment, at least about 99.3% of compounds ofFormula 4 in the composition have the same structure. In one embodiment,at least about 99.5% of compounds of Formula 4 in the composition havethe same structure.

A fourth aspect disclosed herein are high-purity compositions comprisinga compound of Formula 5

wherein R₁ and R₂ are independently H or C₁-C₆ alkyl; each x isindependently selected from the group consisting of 2, 3, 4, 5, and 6;and wherein at least about 98.4% of compounds of Formula 5 in thecomposition have the same structure. In one embodiment, R₁ and R₂ are H.In one embodiment, each x is 3.

In one embodiment, at least about 98.4% of compounds of Formula 5 in thecomposition have the same molecular weight, wherein the same molecularweight excludes isotopic variations. In one embodiment, at least about98.7% of compounds of Formula 5 in the composition have the samemolecular weight, wherein the same molecular weight excludes isotopicvariations. In one embodiment, at least about 99% of compounds ofFormula 5 in the composition have the same molecular weight, wherein thesame molecular weight excludes isotopic variations. In one embodiment,at least about 99.3% of compounds of Formula 5 in the composition havethe same molecular weight, wherein the same molecular weight excludesisotopic variations. In one embodiment, at least about 99.5% ofcompounds of Formula 5 in the composition have the same molecularweight, wherein the same molecular weight excludes isotopic variations.In one embodiment, at least about 98.7% of compounds of Formula 5 in thecomposition have the same structure. In one embodiment, at least about99% of compounds of Formula 5 in the composition have the samestructure. In one embodiment, at least about 99.3% of compounds ofFormula 5 in the composition have the same structure. In one embodiment,at least about 99.5% of compounds of Formula 5 in the composition havethe same structure.

A fifth aspect disclosed herein are methods for synthesizing a purifiedsample of a compound of Formula 4

wherein R₁ and R₂ are independently H or C₁-C₆ alkyl; each x isindependently selected from the group consisting of 2, 3, 4, 5, and 6;and wherein at least about 98.4% of the compounds of Formula 4 in thepurified sample have the same structure comprising: reacting a compoundof Formula 5

with about 90% HNO₃ in AcOH at about 0-5° C. for at least 30 minutes,wherein at least about 98.4% of the compounds of Formula 4 formed in thepurified sample have the same structure. In one embodiment, R₁ and R₂are H. In one embodiment, each x is 3.

In one embodiment, at least about 98.4% of compounds of Formula 4 in thepurified sample have the same molecular weight, wherein the samemolecular weight excludes isotopic variations. In one embodiment, atleast about 98.7% of compounds of Formula 4 in the purified sample havethe same molecular weight, wherein the same molecular weight excludesisotopic variations. In one embodiment, at least about 99% of compoundsof Formula 4 in the purified sample have the same molecular weight,wherein the same molecular weight excludes isotopic variations. In oneembodiment, at least about 99.3% of compounds of Formula 4 in thepurified sample have the same molecular weight, wherein the samemolecular weight excludes isotopic variations. In one embodiment, atleast about 99.5% of compounds of Formula 4 in the purified sample havethe same molecular weight, wherein the same molecular weight excludesisotopic variations. In one embodiment, at least about 98.7% ofcompounds of Formula 4 in the purified sample have the same structure.In one embodiment, at least about 99% of compounds of Formula 4 in thepurified sample have the same structure. In one embodiment, at leastabout 99.3% of compounds of Formula 4 in the purified sample have thesame structure. In one embodiment, at least about 99.5% of compounds ofFormula 4 in the purified sample have the same structure.

A sixth aspect disclosed are methods for synthesizing a purified sampleof a compound of Formula 5

wherein R₁ and R₂ are independently H or C₁-C₆ alkyl; each x isindependently selected from the group consisting of 2, 3, 4, 5, and 6;and wherein at least about 98.4% of the compounds of Formula 5 in thepurified sample have the same structure comprising: reacting tosylatedcompound of Formula 7

with o-dihydroxybenzene of Formula 6

in the presence of K₂CO₃ and an aprotic solvent and heating the mixtureat about 65° C. for about 24 hr wherein at least about 98.4% of thecompound of Formula 5 formed in the purified sample have the samestructure. In a further or alternative embodiment, the mixture is heatedat the reflux temperature of the aprotic solvent.

In one embodiment, aprotic solvent is selected from the group consistingof aldehydes, ketones, dimethyl sulfoxide, dimethyl formamide, pyridine,diethyl ether, ethyl acetate, propyl acetate, isopropyl acetate, butylacetate, acetonitrile, benzonitrile, sulfur dioxide, tetrahydrofuran,and hexamethyl phosphoramide. In one embodiment, the aprotic solvent istetrahydrofuran. In a further or alternative embodiment, R₁ and R₂ areH. In one embodiment, each x is 3, and further, the aprotic solvent istetrahydrofuran.

In one embodiment, at least about 98.4% of compounds of Formula 5 in thepurified sample have the same molecular weight, wherein the samemolecular weight excludes isotopic variations. In one embodiment, atleast about 98.7% of compounds of Formula 5 in the purified sample havethe same molecular weight, wherein the same molecular weight excludesisotopic variations. In one embodiment, at least about 99% of compoundsof Formula 5 in the purified sample have the same molecular weight,wherein the same molecular weight excludes isotopic variations. In oneembodiment, at least about 99.3% of compounds of Formula 5 in thepurified sample have the same molecular weight, wherein the samemolecular weight excludes isotopic variations. In one embodiment, atleast about 99.5% of compounds of Formula 5 in the purified sample havethe same molecular weight, wherein the same molecular weight excludesisotopic variations. In one embodiment, at least about 98.7% ofcompounds of Formula 5 in the purified sample have the same structure.In one embodiment, at least about 99% of compounds of Formula 5 in thepurified sample have the same structure. In one embodiment, at leastabout 99.3% of compounds of Formula 5 in the purified sample have thesame structure. In one embodiment, at least about 99.5% of compounds ofFormula 5 in the purified sample have the same structure.

A seventh aspect disclosed herein are pharmaceutical compositionscomprising a compound according to Formula 1 and a pharmaceuticallyacceptable carrier, wherein at least about 98.4% of the compounds ofFormula 1 in the pharmaceutical composition have the same structure. Inone embodiment, M is Gd⁺³. In one embodiment, R₄ and R₇ are C₃H₆OH; R₅and R₆ are C₂H₅; R₃ and R₈ are CH₃; R₁ and R₂ are H. In one embodiment,each x is 3. In one embodiment, each X is AcO⁻. In another embodiment, Mis Lu⁺³. In a further embodiment, R₄ and R₇ are C₃H₆OH; R₅ and R₆ areC₂H₅; R₃ and R₈ are CH₃; R₁ and R₂ are H. In a further embodiment, eachx is 3. In a further embodiment, each X is AcO⁻. In a further oralternate embodiment, each X is selected from the group consisting ofsugar derivatives, cholesterol derivatives, PEG acids, organic acids,organosulfates, organophosphates, phosphates or inorganic ligands. In afurther or alternate embodiment, X is derived from an acid selected fromthe group consisting of gluconic acid, glucoronic acid, cholic acid,deoxycholic acid, methylphosphonic acid, phenylphosphonic acid,phosphoric acid, formic acid, propionic acid, butyric acid, pentanoicacid, 3,6,9-trioxodecanoic acid, 3,6-dioxoheptanoic acid,2,5-dioxoheptanoic acid, methylvaleric acid, glycolic acid, pyruvicacid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid,fumaric acid, tartaric acid, citric acid, methanesulfonic acid,ethanesulfonic acid, benzoic acid, salicylic acid, 3-fluorobenzoic acid,4-aminobenzoic acid, cinnamic acid, mandelic acid, andp-toluene-sulfonic acid.

In one embodiment, at least about 98.4% of compounds of Formula 1 in thepharmaceutical composition have the same molecular weight, wherein thesame molecular weight excludes isotopic variations. In one embodiment,at least about 98.7% of compounds of Formula 1 in the pharmaceuticalcomposition have the same molecular weight, wherein the same molecularweight excludes isotopic variations. In one embodiment, at least about99% of compounds of Formula 1 in the pharmaceutical composition have thesame molecular weight, wherein the same molecular weight excludesisotopic variations. In one embodiment, at least about 99.3% ofcompounds of Formula 1 in the pharmaceutical composition have the samemolecular weight, wherein the same molecular weight excludes isotopicvariations. In one embodiment, at least about 99.5% of compounds ofFormula 1 in the pharmaceutical composition have the same molecularweight, wherein the same molecular weight excludes isotopic variations.In one embodiment, at least about 98.7% of compounds of Formula 1 in thepharmaceutical composition have the same structure. In one embodiment,at least about 99% of compounds of Formula 1 in the pharmaceuticalcomposition have the same structure. In one embodiment, at least about99.3% of compounds of Formula 1 in the pharmaceutical composition havethe same structure. In one embodiment, at least about 99.5% of compoundsof Formula 1 in the pharmaceutical composition have the same structure.In one embodiment, the pharmaceutically acceptable carrier is suitablefor parenteral administration.

An eighth aspect disclosed herein, involves a method of treating acancer by administering to a subject in need thereof an effective amountof a purified composition comprising a compound of Formula 1 wherein atleast about 98.4% of the compounds of Formula 1 in the purifiedcomposition have the same structure, or a pharmaceutically acceptablesalt, solvate, isomers, tautomers, metabolites, analogs, or prodrugsthereof. In one embodiment, the method further comprises of surgery,radiation therapy, chemotherapy, gene therapy, immunotherapy, or acombination thereof. In one embodiment, the cancer is selected from thegroup consisting of glioblastoma, lymphomas, leukemia, renal cellcancer, head and neck, cancer, breast cancer, prostrate cancer, ovariancancer, and lung, cancer.

A ninth aspect disclosed herein, involves a method of treating acardiovascular disease by administering to a subject in need thereof aneffective amount of a purified composition comprising a compound ofFormula 1 wherein at least about 98.4% of the compounds of Formula 1 inthe purified composition have the same structure,

or a pharmaceutically acceptable salt, solvate, isomers, tautomers,metabolites, analogs, or prodrugs thereof. In one embodiment, thecardiovascular disease is selected from the group consisting of coronaryartery disease, saphenous vein graft disease, and peripheral arterydisease. In a further embodiment, M is Lu⁺³ and the subject is furtherprovided photodynamic therapy. In one embodiment, the photodynamictherapy is provided after the subject has been administered the compoundof Formula 1. In another embodiment, the target area is illuminated withlight having wavelengths between about 725 and 760 nm. In a further oralternate embodiment, each X is selected from the group consisting ofsugar derivatives, cholesterol derivatives, PEG acids, organic acids,organosulfates, organophosphates, phosphates or inorganic ligands. In afurther or alternate embodiment, X is derived from an acid selected fromthe group consisting of gluconic acid, glucoronic acid, cholic acid,deoxycholic acid, methylphosphonic acid, phenylphosphonic acid,phosphoric acid, formic acid, propionic acid, butyric acid, pentanoicacid, 3,6,9-trioxodecanoic acid, 3,6-dioxoheptanoic acid,2,5-dioxoheptanoic acid, methylvaleric acid, glycolic acid, pyruvicacid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid,fumaric acid, tartaric acid, citric acid, methanesulfonic acid,ethanesulfonic acid, benzoic acid, salicylic acid, 3-fluorobenzoic acid,4-aminobenzoic acid, cinnamic acid, mandelic acid, andp-toluene-sulfonic acid.

In one embodiment of the aforementioned eighth and ninth aspects, M isGd⁺³. In one embodiment, R₄ and R₇ are C₃H₆OH; R₅ and R₆ are C₂H₅; R₃and R₈ are CH₃; R₁ and R₂ are H. In one embodiment, each x is 3. In oneembodiment, each X is AcO⁻. In another embodiment, M is Lu⁺³. In afurther embodiment, R₄ and R₇ are C₃H₆OH; R₅ and R₆ are C₂H₅; R₃ and R₈are CH₃; R₁ and R₂ are H. In a further embodiment, each x is 3. In afurther embodiment, each X is AcO⁻. In a further or alternateembodiment, each X is selected from the group consisting of sugarderivatives, cholesterol derivatives, PEG acids, organic acids,organosulfates, organophosphates, phosphates or inorganic ligands. In afurther or alternate embodiment, X is derived from an acid selected fromthe group consisting of gluconic acid, glucoronic acid, cholic acid,deoxycholic acid, methylphosphonic acid, phenylphosphonic acid,phosphoric acid, formic acid, propionic acid, butyric acid, pentanoicacid, 3,6,9-trioxodecanoic acid, 3,6-dioxoheptanoic acid,2,5-dioxoheptanoic acid, methylvaleric acid, glycolic acid, pyruvicacid, oxalic acid, malic acid, malonic acid, succinic acid, maleic acid,fumaric acid, tartaric acid, citric acid, methanesulfonic acid,ethanesulfonic acid, benzoic acid, salicylic acid, 3-fluorobenzoic acid,4-aminobenzoic acid, cinnamic acid, mandelic acid, andp-toluene-sulfonic acid.

In one embodiment of the aforementioned eighth and ninth aspects, atleast about 98.4% of compounds of Formula 1 in the composition have thesame molecular weight, wherein the same molecular weight excludesisotopic variations. In one embodiment, at least about 98.7% ofcompounds of Formula 1 in the composition have the same molecularweight, wherein the same molecular weight excludes isotopic variations.In one embodiment, at least about 99% of compounds of Formula 1 in thecomposition have the same molecular weight, wherein the same molecularweight excludes isotopic variations. In one embodiment, at least about99.3% of compounds of Formula 1 in the composition have the samemolecular weight, wherein the same molecular weight excludes isotopicvariations. In one embodiment, at least about 99.5% of compounds ofFormula 1 in the composition have the same molecular weight, wherein thesame molecular weight excludes isotopic variations. In one embodiment,at least about 98.7% of compounds of Formula 1 in the composition havethe same structure. In one embodiment, at least about 99% of compoundsof Formula 1 in the composition have the same structure. In oneembodiment, at least about 99.3% of compounds of Formula 1 in thecomposition have the same structure. In one embodiment, at least about99.5% of compounds of Formula 1 in the composition have the samestructure.

The term “chemotherapy” as described herein, refers to theadministration of one or more anti-cancer drugs and/or other agents to acancer patient by various methods, including intravenous, oral,intramuscular, intraperitoneal, intravesical, subcutaneous, transdermal,buccal, or inhalation or in the form of a suppository.

The term “old process” as described herein, refers to process describedin Sessler et al in U.S. Pat. No. 5,994,535, incorporated herein byreference in its entirety.

The term “purified sample” or “purified composition” refers to acomposition having a high degree of uniformity in the chemical structure(or molecular weight, excluding isotopic variations) of compounds havinga particular generic chemical formula. That is, multiple compounds in acomposition may be species of a particular generic chemical formula;however in a purified sample or purified composition, a high proportionof such compounds have the same chemical formula (or molecular weight,excluding isotopic variations). The value for the terms “high degree” or“high proportion” are provided specific values herein, for example, atleast about 98.4%, at least about 98.7%, and the like. A purified sampleor purified composition can include other chemical entities, includingsolvents, salts, reagents, pharmaceutical excipients, additionaltherapeutic agents, and the like. The defining aspect is that thecomposition has a high degree of uniformity in the chemical structure(or molecular weight, excluding isotopic variations) of compounds havinga particular generic chemical formula (or molecular weight, excludingisotopic variations), and in particular compounds having the structureof Formula 1, Formula 2, Formula 4 and Formula 5. The uniformity of aparticular purified composition or purified sample can be determined bya variety of analytical methods, including by way of example only,chromatography, and spectroscopic or spectrometric methods.

The term “parenteral administration” as described herein, refers toadministration of at least one agent by means other than through thealimentary tract. Parenteral routes of administration involve injectionsinto various compartments of the body such as but not limited to,intravenous, subcutaneous, intramuscular, intraperitoneal and the like.

The terms “pharmaceutically effective amount” or “effective amount” asdescribed herein, refers to a nontoxic but sufficient amount of theagent to provide the desired biological, therapeutic, and/orprophylactic result. The desired results include reduction and/oralleviation of the signs, symptoms, or causes of a disease, or any otherdesired alteration of a biological system. An effective amount in anyindividual case may be determined by one of ordinary skill in the artusing routine experimentation.

The term “pharmaceutically acceptable” or “pharmacologically acceptable”as described herein, may mean a material which is not biologically orotherwise undesirable, i.e., the material may be administered to anindividual without causing any undesirable biological effects orinteracting in a deleterious manner with any of the components of thecomposition in which it is contained.

The term “photodynamic therapy” as described herein, refers to atreatment that combines a light source and a photosensitizing agent (adrug that is activated by light).

The term “radiation therapy” as described herein, refers to exposing apatient to high-energy radiation, including without limitation x-rays,gamma rays, and neutrons. This type of therapy includes withoutlimitation external-beam therapy, internal radiation therapy, implantradiation, brachytherapy, systemic radiation therapy, and radiotherapy.

The term “same molecular weight,” as used herein, refers to compoundsthat have the same molecular weight, excluding isotopic variations andthe identity of the counterions. That is, the molecular weight of acompound is determined by adding up the atomic weight of all atoms inthe formula, excluding however, the counterions (i.e., the X groups inFormula 1). In particular, a compound does not have a differentmolecular weight from another compound, for purposes of this definitionbecause it has a 2H instead of a 1H in a structure (i.e., isotopicvariations do not constitute different molecular weights). By way ofexample only, compounds having the same molecular weight in reference toFormula 1 have the same M, R3, R4, R5, R6, R7, R8, R1, and R2 groups,and the same values for n, y, and x; however, the identity of the Xgroup or groups can be different.

The term “same structure,” as used herein, refers to compounds havingidentical chemical structures, excluding isotopic variations andcounterions. By way of example only, compounds having the same structurein reference to Formula 1 have the same M, R3, R4, R5, R6, R7, R8, R1,and R2 groups, and the same values for n, y, and x; however, theidentity of the X group or groups can be different.

The term “surgery” as described herein, refers to any therapeutic ordiagnostic procedure that involves methodical action of the hand or ofthe hand with an instrument, on the body of a human or other mammal, toproduce a curative, remedial, or diagnostic effect.

The term “treating” and its grammatical equivalents as described herein,refers to achieving, or attempting to achieve, a therapeutic benefitand/or a prophylactic benefit. By therapeutic benefit is meanteradication or amelioration, at least in part, of the underlyingdisorder being treated. For example, in a cancer patient, therapeuticbenefit includes eradication or amelioration, at least in part, of theunderlying cancer. Also, a therapeutic benefit includes the eradicationor amelioration, at least in part, of one or more of the physiologicalsymptoms associated with the underlying disorder such that animprovement is observed in the patient, notwithstanding the fact thatthe patient may still be afflicted with the underlying disorder. Forprophylactic benefit, a method disclosed herein may be performed on, ora composition disclosed herein administered to, a patient at risk ofdeveloping cancer, or to a patient reporting one or more of thephysiological symptoms of such conditions, even in the absence of adiagnosis of the condition.

INCORPORATION BY REFERENCE

Unless stated otherwise, all publications and patent applicationsmentioned in this specification are herein incorporated by reference tothe same extent as if each individual publication or patent applicationwas specifically and individually indicated to be incorporated byreference.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1 schematically summarizes an illustrative synthesis of texaphyrinmetal complexes described herein.

FIG. 2 shows an illustrative chromatogram comparing the purity of acompound of Formula 4 as synthesized by the old process and the improvedprocess disclosed herein.

FIG. 3 shows an illustrative chromatogram comparing the purity of acompound of Formula 1 as synthesized by the old process and the improvedprocess disclosed herein.

DETAILED DESCRIPTION OF THE INVENTION Texaphyrins

Sessler et al reported in U.S. Pat. No. 5,994,535, incorporated hereinby reference in its entirety, the synthesis of “texaphyrins”, whichabsorb strongly in the tissue-transparent 730-770 nm range. Thiscompound is capable of existing in both its free-base form and ofsupporting the formation of complexes with a variety of metal cations,such as Cd²⁺, Hg²⁺, In³⁺, Y³⁺, Nd³⁺, Eu³⁺, Sm³⁺, La³⁺, Lu³⁺, Gd³⁺, andother cations of the lanthanide series. Texaphyrins find use in thetreatment of cancer and atherosclerosis.

Compositions of texaphyrins synthesized by the old process werepolydisperse at the polyethylene glycol chain length, that is,compositions of texaphyrins synthesized by the old process comprisedmultiple species that differed in structure, although sharing a commongeneric chemical structure. This variety in structure arose, at least inpart, from non-uniformity in the identity of the polyethylene glycolsubstituents. As a result, high purity samples (i.e., having less thanabout 1.6% polydispersity at the polyethylene glycol chain) were notavailable for therapeutic use. The synthetic processes disclosed herein,provides a breakthrough in this area by providing a method for thesynthesis of high purity texaphyrins (i.e., having less than about 1.6%polydispersity at the polyethylene glycol chain).

There are a number of disadvantages associated with low purity in asample, including, by way of example only, difficulties in solubilityoptimization in pharmaceutical formulations such that the effectiveconcentrations of an agent may vary from preparation to preparation;difficulties in optimizing drug dosage; and difficulties in stabilizingformulations. The improvement in the purities of the compounds disclosedherein overcome such disadvantages of solubility, dose optimization, anddrug stabilization. The improvement in the purity may further result ina decrease in side effects associated with low purity pharmaceuticalformulations, an improvement in the measured half life of the drugformulation, and an improvement in drug targeting.

Representative Synthesis of High-Purity Samples or Compositions

Without limiting the scope of the compositions and methods describedherein, a representative synthesis of texaphyrin metal complexes isdepicted in FIG. 1. A compound of Formula 8 (each x is 3) is tosylatedto a compound of Formula 7. The tosylated compound of Formula 7 istreated with a o-dihydroxybenzene derivative of Formula 6 in thepresence of K₂CO₃ and an aprotic solvent to result in a compound ofFormula 5. The aprotic solvent, by way of example only, may be selectedfrom the group consisting of aldehydes, ketones, dimethyl sulfoxide,dimethyl formamide, pyridine, diethyl ether, ethyl acetate, propylacetate, isopropyl acetate, butyl acetate, acetonitrile, benzonitrile,sulfur dioxide, tetrahydrofuran, hexamethyl phosphoramide, and the like.In one embodiment disclosed herein, the solvent is tetrahydrofuran.

The compound of Formula 5 is nitrated in the presence of nitric acid andacetic acid to result in a compound of Formula 4. A high purity sampleof a compound having the structure of Formula 4 is thus obtained,wherein at least about 98.4% of the compounds of Formula 4 in the highpurity sample have the same structure (and the same molecular weight,excluding isotopic variation), i.e. both polyethylene glycol chainlengths on the aromatic moiety have the same chain length.

In some embodiments disclosed herein, at least about 98.7%, 99%, 99.3%or 99.5% of the compounds of Formula 4 in the high purity sample havethe same structure (and the same molecular weight, excluding isotopicvariation), i.e. both polyethylene glycol chain lengths on the aromaticmoiety have the same chain length (including the same “x” value per thegeneric formula).

Table 1 compares the purity of compositions comprising compounds ofFormula 4 made via the old process and the process disclosed herein.FIG. 2 is a chromatogram comparison of the purity of compositionscomprising compounds of Formula 4 made via the old process and theprocess disclosed herein.

TABLE 1 Purity of compositions comprising compounds of Formula 4 BatchNumber Purity PEG impurities Old Process Batch 1 97.8% 1.96% Batch 298.2% 1.61% Batch 3 98.2% 1.63% Batch 4 97.7% 2.01% New Process Batch 199.6% 0.23% Batch 2 99.6% 0.21% Batch 3 99.4% 0.28% Batch 4 99.4% 0.24%

The higher purity of the compounds may be attributable to a solvent usedin the formation of the compound of Formula 5. During development of thecompound of Formula 4, one particular impurity was identified as havinga structure shown below:

This impurity was found in greater quantity than would be expected basedon an amount of ethylene glycol monomethyl ether contained in compoundof Formula 8. Without limiting the scope of the compositions and methodsdescribed herein, it is postulated that this impurity originates in thereaction step of the synthesis of compound of Formula 5 from compound ofFormula 7 via an intermediate A shown below:

The 1,4-dioxane type molecule A is formed by an intramolecular attack ofan ether oxygen to displace tosylate. The o-dihydroxybenzene derivativeof Formula 6 can react with A to form either compound of Formula 5 orthe monoethylene glycol/triethylene glycol impurity (mono/tri) as shown:

Without limiting the scope of the compositions and methods describedherein, it is postulated that the 1,4-dioxane cation A might bestabilized in the ethanol solvent used in the old process. In methodsdisclosed herein, THF minimizes the formation of the mono/tri impurity.Table 2 depicts the % of mono/tri impurity obtained in differentsolvents. Ethanol results in 0.43% of mono/tri impurity whereas THFgives only 0.08% of mono/tri impurity.

TABLE 2 Comparison of Mono/tri impurity in different solvents Compoundof Formula 4 (solids) Compound of Formula 5 Yield Conv. % Com- % (fromo- Time pound of mono/tri dihydroxy Solvent Temperature (hr) Formula 4impurity benzene) Ethanol 78° C. 6 99 0.43 68% Ethanol 50° C. >48 n/an/a n/a Acetonitrile 80° C. 8 98.6 0.41 79% MTBE 58-60° C. 15 93 4.6 ~50%  Propyl 90° C. 2 97.9 0.13 66% acetate THF 70° C. 4 99.2 0.08 47%

The compound of Formula 4 after reduction results in a phenylenediamineof Formula 3a which on condensation with a compound of Formula 3bresults in a high purity sample, wherein at least about 98.4% of thecompounds having a structure of Formula 2 in the high purity sample havethe same structure (and the same molecular weight, excluding isotopicvariation), i.e. both polyethylene glycol chain lengths on the aromaticmoiety have the same chain length. In some embodiments disclosed herein,at least about 98.7%, 99%, 99.3% or 99.5% of the compounds of Formula 2in the high purity sample have the same structure (and the samemolecular weight, excluding isotopic variation), i.e. both polyethyleneglycol chain lengths on the aromatic moiety have the same chain length(including the same “x” value per the generic formula).

The compound of Formula 2 complexes oxidatively with a trivalent metalcation (M) to result in a high purity sample, wherein at least about98.4% of the compounds having a structure of Formula 1 in the highpurity sample have the same structure (and the same molecular weight,excluding isotopic variation), i.e. both polyethylene glycol chainlengths on the aromatic moiety have the same chain length. In someembodiments disclosed herein, at least about 98.7%, 99%, 99.3% or 99.5%of the compounds of Formula 1 in the high purity sample have the samestructure (and the same molecular weight, excluding isotopic variation),i.e. both polyethylene glycol chain lengths on the aromatic moiety havethe same chain length (including the same “x” value per the genericformula). Suitable M include, by way of example only, Mn⁺³, Co⁺³, Ni⁺³,Y⁺³, In⁺³, Pr⁺³, Nd⁺³, Sm⁺³, Eu⁺³, Gd⁺³, Tb⁺³, Dy⁺³, Er⁺³, Fe⁺³, Ho⁺³,Ce⁺³, Tm⁺³, Yb⁺³, Lu⁺³, La⁺³, and U⁺³; further examples of M includedivalent metal cations, such as Mn⁺², Co⁺², Ni⁺², and Fe⁺². In someembodiments disclosed herein, the trivalent metal ions are Gd⁺³ or Lu⁺³.Table 3 compares the purities of samples of compounds of Formula 1 madeby the old process and in the process disclosed herein. The old processresulted in 96.6% purity whereas the process disclosed herein improvesit to at least about 98.4%. FIG. 3 is a chromatogram comparison of thepurity of compositions comprising compounds of Formula 1 made via theold process and the process disclosed herein.

TABLE 3 Purity of compositions comprising compounds of Formula 1 BatchNumber Purity PEG impurities Old Process Batch 1 96.0% 2.45% Batch 295.7% 2.66% Batch 3 96.5% 2.44% Batch 4 96.6% 2.32% Batch 5 95.2% 2.74%New Process Batch 1 99.3% 0.33% Batch 2 99.1% 0.38% Batch 3 99.2% 0.33%Batch 4 99.2% 0.25% Batch 5 99.0% 0.30%

In the condensation step between compound 3a and 3b, the introduction ofvarious substituents on the tripyrrole portion of the molecule may beaccomplished by appropriate substitutions in the 3 and/or 4 positions ofthe pyrrole rings at a synthetic step prior to condensation with thesubstituted phenylenediamine. The substituents on the benzene ring maybe obtained by appropriate substitutions in the benzene ring prior tocondensation with the pyrrole rings.

Illustrative Methods for Determining Purity of a Sample or Composition

Without limiting the scope of the compositions and methods disclosedherein, some of the methods for demonstrating a purity of a compound aredescribed.

Chromatographic methods that are contemplated to be used fordemonstrating purity are, by way of example only, molecular sizeexclusion chromatography, native gel electrophoresis, high pressureliquid chromatography (HPLC), liquid chromatography (LC), liquidchromatography coupled with mass spectroscopy (LC/MS), gaschromatography (GC), GC coupled with mass spectroscopy (GC MS),supercritical fluid chromatography, gel permeation chromatography andion exchange chromatography, and Reversed-Phase High Performance LiquidChromatography.

Other methods that are contemplated to be used for demonstrating purityare, by way of example only, end group analysis, vapor pressureosmometry, cryoscopy/ebulliometry (freezing point depression/boilingpoint elevation), viscometry, small-angle X ray scattering, laser lightscattering, optical absorption and scattering, ultracentrifugation,field flow fractionation, matrix-assisted laser desorption/ionizationtime-of-flight (MALDI-TOF) mass spectrometry, nuclear magnetic resonancespectrometry and crystallization.

Method for Treating Cancer

Without limiting the scope of the compositions and the methods disclosedherein, the methods are used to treat several specific cancers ortumors. Cancer types include (some of which may overlap in scope), byway of example only, adrenal cortical cancer, anal cancer, aplasticanemia, bile duct cancer, bladder cancer, bone cancer, bone metastasis,adult CNS brain tumors, pediatric CNS brain metastases, brainmetastases, breast cancer, Castleman Disease, cervical cancer, childhoodNon-Hodgkin's lymphoma, colon and rectum cancer, endometrial cancer,esophagus cancer, Ewing's family of tumors, eye cancer, gallbladdercancer, gastrointestinal carcinoid tumors, gastrointestinal stromaltumors, gestational trophoblastic disease, hematological malignancies,Hodgkin's disease, Kaposi' sarcoma, kidney cancer, laryngeal andhypopharyngeal cancer, acute lymphocytic leukemia, acute myeloidleukemia, children's leukemia, chronic lymphocytic leukemia, chronicmyeloid leukemia, liver cancer, lung cancer, lung carcinoid tumors,Non-Hodgkin's lymphoma, male breast cancer, malignant mesothelioma,multiple myeloma, myelodysplastic syndrome, nasal cavity and paranasalcancer, nasopharyngeal cancer, neuroblastoma, oral cavity andoropharyngeal cancer, osteosarcoma, ovarian cancer, pancreatic cancer,penile cancer, pituitary tumor, prostate cancer, retinoblastoma,rhabdomyosarcoma, salivary gland cancer, sarcoma (adult soft tissuecancer), melanoma skin cancer, nonmelanoma skin cancer, stomach cancer,testicular cancer, thymus cancer, thyroid cancer, uterine sacrcoma,vaginal cancer, vulvar cancer, and Waldenstrom's macroglobulinemia. Inone embodiment, the cancers are selected from the group consisting ofmetastatic brain cancer, lung cancer, glioblastoma, lymphomas, leukemia,renal cell cancer (kidney cancer), head and neck cancer, breast cancer,prostrate cancer, and ovarian cancer.

Disclosed herein are methods and compositions to treat lung cancercomprising administration of the high-purity compositions of Formula 1.Treatment options for lung cancer include (which can be provided to apatient in conjunction with administration of the high-puritycompositions of Formula 1), by way of example only, surgery,immunotherapy, radiation therapy, chemotherapy, photodynamic therapy, ora combination thereof. Some possible surgical options for treatment oflung cancer are a segmental or wedge resection, a lobectomy, or apneumonectomy. Radiation therapy may be external beam radiation therapyor brachytherapy.

Disclosed herein are methods and compositions to treat CNS neoplasmscomprising administration of the high-purity compositions of Formula 1.Treatment options for CNS neoplasms include (which can be provided to apatient in conjunction with administration of the high-puritycompositions of Formula 1), by way of example only, surgery, radiationtherapy, immunotherapy, hyperthermia, gene therapy, chemotherapy, andcombination of radiation and chemotherapy. Doctors also may prescribesteroids to reduce the swelling inside the CNS.

Disclosed herein are methods to treat kidney cancer comprisingadministration of the high-purity compositions of Formula 1. Kidneycancer (also called renal cell cancer or renal adenocarcinoma) is adisease in which malignant cells are found in the lining of tubules inthe kidney. Treatment options for kidney cancer include (which can beprovided to a patient in conjunction with administration of thehigh-purity compositions of Formula 1), by way of example only, surgery,radiation therapy, chemotherapy and immunotherapy. Some possiblesurgical options to treat kidney cancer include, by way of example only,partial nephrectomy, simple nephrectomy and radical nephrectomy.Radiation therapy may be external beam radiation therapy orbrachytherapy. Stem cell transplant may be used to treat kidney cancer.

In one embodiment disclosed herein are methods to treat lymphomacomprising administration of the high-purity compositions of Formula 1.Treatment options for lymphoma include (which can be provided to apatient in conjunction with administration of the high-puritycompositions of Formula 1), by way of example only, chemotherapy,immunotherapy, radiation therapy and high-dose chemotherapy with stemcell transplant. Radiation therapy may be external beam radiationtherapy or brachytherapy.

Disclosed herein are methods for treating breast cancer comprisingadministration of the high-purity compositions of Formula 1. Treatmentoptions for breast cancer include (which can be provided to a patient inconjunction with administration of the high-purity compositions ofFormula 1), by way of example only, surgery, immunotherapy, radiationtherapy, chemotherapy, endocrine therapy, or a combination thereof. Alumpectomy and a mastectomy are two possible surgical proceduresavailable for breast cancer patients.

Disclosed herein are methods for treating ovarian cancer, comprisingadministration of the high-purity compositions of Formula 1. Treatmentoptions for ovarian cancer include (which can be provided to a patientin conjunction with administration of the high-purity compositions ofFormula 1), by way of example only, surgery, immunotherapy,chemotherapy, hormone therapy, radiation therapy, or combinationsthereof. Some possible surgical procedures include debulking, and aunilateral or bilateral oophorectomy and/or a unilateral or bilateralsalpigectomy.

Disclosed herein are methods for treating cervical cancer, comprisingadministration of the high-purity compositions of Formula 1. Treatmentoptions for cervical cancer include (which can be provided to a patientin conjunction with administration of the high-purity compositions ofFormula 1), by way of example only, surgery, immunotherapy, radiationtherapy and chemotherapy. Some possible surgical options arecryosurgery, a hysterectomy, and a radical hysterectomy. Radiationtherapy for cervical cancer patients includes external beam radiationtherapy or brachytherapy.

Disclosed herein are methods to treat prostate cancer, comprisingadministration of the high-purity compositions of Formula 1. Treatmentoptions for prostate cancer include (which can be provided to a patientin conjunction with administration of the high-purity compositions ofFormula 1), by way of example only, surgery, immunotherapy, radiationtherapy, cryosurgery, hormone therapy, and chemotherapy. Possiblesurgical procedures to treat prostate cancer include, by way of exampleonly, radical retropubic prostatectomy, a radical perinealprostatectomy, and a laparoscopic radical prostatectomy. Some radiationtherapy options are external beam radiation, including three dimensionalconformal radiation therapy, intensity modulated radiation therapy, andconformal proton beam radiation therapy. Brachytherapy (seedimplantation or interstitial radiation therapy) is also an availablemethod of treatment for prostate cancer. Cryosurgery is another possiblemethod used to treat localized prostate cancer cells. Hormone therapy,also called androgen deprivation therapy or androgen suppressiontherapy, may be used to treat prostate cancer. Several methods of thistherapy are available including an orchiectomy in which the testicles,where 90% of androgens are produced, are removed. Another method is theadministration of luteinizing hormone-releasing hormone (LHRH) analogsto lower androgen levels. The LHRH analogs available include leuprolide,nafarelin, goserelin, triptorelin, and histrelin. An LHRH antagonist mayalso be administered, such as abarelix. Treatment with an antiandrogenagent, which blocks androgen activity in the body, is another availabletherapy. Such agents include flutamide, bicalutamide, and nilutamide.This therapy is typically combined with LHRH analog administration or anorchiectomy, which is termed a combined androgen blockade (CAB).Chemotherapy may be appropriate where a prostate tumor has spreadoutside the prostate gland and hormone treatment is not effective.Anti-cancer drugs may be administered to slow the growth of prostatecancer, reduce symptoms and improve the quality of life.

Disclosed herein are methods for treating leukemia, comprisingadministration of the high-purity compositions of Formula 1. Treatmentoptions for leukemia include (which can be provided to a patient inconjunction with administration of the high-purity compositions ofFormula 1), by way of example only, immunotherapy, radiation therapy,chemotherapy, bone marrow or peripheral blood stem cell transplantation,or a combination thereof. Radiation therapy includes external beamradiation and may have side effects. Anti-cancer drugs may be used inchemotherapy to treat leukemia. Monoclonal antibody therapy may be usedto treat AML patients. Small molecules or radioactive chemicals may beattached to these antibodies before administration to a patient in orderto provide a means of killing leukemia cells in the body. The monoclonalantibody, gemtuzumab ozogamicin, which binds CD33 on AML cells, may beused to treat AML patients unable to tolerate prior chemotherapyregimens. Bone marrow or peripheral blood stem cell transplantation maybe used to treat AML patients. Some possible transplantation proceduresare an allogenic or an autologous transplant.

Disclosed herein are methods and compositions to treat head and neckcancer, comprising administration of the high-purity compositions ofFormula 1. Treatment options for head and neck cancer include (which canbe provided to a patient in conjunction with administration of thehigh-purity compositions of Formula 1), by way of example only, surgery,radiation, chemotherapy, combined modality therapy, gene therapy, eitheralone or in combination thereof.

Method for Treating Cardiovascular Disease

Disclosed herein are the methods to treat cardiovascular diseases.Cardiovascular disease has two main components: Diseases of the heart(cardio) and Diseases of the blood vessels (vascular).

Coronary Artery Disease

These are diseases of the arteries that supply the heart muscle withblood. Sometimes known as CAD, coronary artery disease is the mostcommon form of heart disease in industrialized nations and far and awaythe leading cause of heart attacks. Coronary artery disease generallymeans that blood flow through the arteries has become impaired. The mostcommon way such obstructions develop is through a condition calledatherosclerosis, a largely preventable type of vascular disease. Thesearteries, whose inner lining is normally smooth, can slowly becomeclogged with clumps of fats, cholesterol and other material, calledatherosclerotic plaques. As a result, the supply of blood—with itsoxygen and nutrients—going to the heart muscle is choked off (myocardialischemia). Chest pain (angina pectoris) occurs, for instance, when theoxygen demand of the heart muscle exceeds the oxygen supply because ofthat narrowing in the coronary arteries. When the imbalance of oxygensupply lasts for more than a few minutes, heart muscle can begin to die,causing a heart attack (myocardial infarction). This may occur withoutsymptoms (silent heart attack), especially in people with diabetes. Inaddition, the lack of blood, even briefly, can lead to serious disordersof the heart rhythm, known as arrhythmias or dysrhythmias. Coronaryartery disease can even cause sudden death from an arrhythmia withoutany prior warning. A heart attack, for instance, can lead to congestiveheart failure, and both of these conditions are types of cardiovasculardisease. Other cardiovascular diseases are, by way of example only,cardiomyopathy, valvular heart disease, pericardial disease, congenitalheart disease, and congestive heart failure.

Diseases of the blood vessels include, by way of example only, highblood pressure, aneurysms, occlusive artery disease, vasculitis, venusthrombosis.

Peripheral Artery Disease (PAD)

In PAD, fatty deposits build up in the inner linings of the arterywalls. These blockages restrict blood circulation, mainly in arteriesleading to the kidneys, stomach, arms, legs and feet. In its earlystages a common symptom is cramping or fatigue in the legs and buttocksduring activity. Such cramping subsides when the person stands still.This is called “intermittent claudication.” People with PAD often havefatty buildup in the arteries of the heart and brain. Because of thisassociation, people with PAD have a higher risk of death from heartattack and stroke. Treatments include, by way of example only, medicinesto help improve walking distance, antiplatelet agents, andcholesterol-lowering agents (statins). Angioplasty or surgery may benecessary.

Saphenous Vein Graft Disease

This term refers to the narrowing, either localized or diffuse, of thesegment of the saphenous vein that has been used as a bypass graft.Serial follow-up studies of patients who have undergone bypass surgeryhave shown that up to 10 percent of vein grafts are occluded by the timeof hospital discharge, which increases to 20 percent by the end of thefirst year after surgery. With time, there is continued development ofdisease and progression, so that by the end of 10 years only one-thirdof the vein grafts that were not occluded at one year are free ofsignificant disease. A re-operation becomes necessary in about 20percent of patients by 10 years. Treatments include, by way of exampleonly, medical therapy, surgery, and angioplasty.

The methods disclosed herein can provide a beneficial effect forpatients suffering from aforementioned diseases, by administration of acompound of Formula 1 or a combination of administration of a compoundof Formula 1 and at least one treatment identified in the foregoingparagraphs, including, by way of example only: surgery, radiationtherapy, chemotherapy, transplantation therapy or the like. The choiceof therapy that can be co-administered with the compositions disclosedherein will depend, in part, on the condition being treated. Forexample, for treating cancer, a compound of Formula 1 can be used incombination with radiation therapy, monoclonal antibody therapy,chemotherapy, bone marrow transplantation, gene therapy, immunotherapy,or a combination thereof. As another example, for treating CAD or PAD, acompound of Formula 1 can be used in combination with surgery, radiationtherapy, chemotherapy, or transplantation therapy.

Formulations, Routes of Administration, and Effective Doses

Another aspect disclosed herein relates to pharmaceutical compositionscomprising a compound of Formula 1 and a pharmaceutically acceptableexcipient. Such pharmaceutical compositions can be used to treat canceror cardiovascular disease in the methods as described herein.

The compounds of Formula 1 may be provided as a prodrug and/or may beallowed to interconvert to compound of Formula 1 in vivo afteradministration. The compounds of Formula 1 and/or its prodrug, or itspharmaceutically acceptable salts may be used in developing aformulation for use in the methods disclosed herein. Further, in someembodiments, the compound may be used in combination with one or moreother compounds or in one or more other forms. The compound of Formula 1may be formulated, in the same dosage unit e.g. in one cream,intravenously-suitable formulation, solution, suppository, tablet, orcapsule.

The term “pharmaceutically acceptable salt” means those salts whichretain the biological effectiveness and properties of the compoundsdisclosed herein, and which are not biologically or otherwiseundesirable. For example, a pharmaceutically acceptable salt does notinterfere with the beneficial effect of the compound disclosed herein intreating a cancer.

Typical salts include by way of example only, salts formed by mixing acompound of Formula 1 in an appropriate buffer, such as phosphatebuffer, or by passing a compound of Formula 1 through an appropriateion-exchange column. In addition, if the compounds disclosed hereincontain a carboxy group or other acidic group, it may be converted intoa pharmaceutically acceptable addition salt with inorganic or organicbases. Examples of suitable bases include by way of example only, sodiumhydroxide, potassium hydroxide, ammonia, tromethamine, meglumine,cyclohexylamine, dicyclohexyl-amine, ethanolamine, diethanolamine andtriethanolamine.

The high-purity compositions of Formula 1 may be administered in eithersingle or multiple doses by any of the accepted modes of administrationof agents having similar utilities, for example as described in thosepatents and patent applications incorporated by reference above,including rectal, buccal, intranasal and transdermal routes, byintra-arterial injection, intravenously, intraperitoneally,parenterally, intramuscularly, subcutaneously, orally, topically, as aninhalant, or via an impregnated or coated device such as a stent, forexample, or an artery-inserted cylindrical polymer.

One mode for administration is parenteral, including, by way of example,by injection. The forms in which the high-purity compositions of Formula1 may be incorporated for administration by injection include aqueous oroil suspensions, or emulsions, with sesame oil, corn oil, cottonseedoil, or peanut oil, as well as elixirs, mannitol, dextrose, or a sterileaqueous solution, and similar pharmaceutical vehicles. Aqueous solutionsin saline are also conventionally used for injection. Ethanol, glycerol,propylene glycol, liquid polyethylene glycol, and the like (and suitablemixtures thereof), cyclodextrin derivatives, and vegetable oils may alsobe employed. The proper fluidity can be maintained, for example, by theuse of a coating, such as lecithin, by the maintenance of the requiredparticle size in the case of dispersion and by the use of surfactants.The prevention of the action of microorganisms can be brought about byvarious antibacterial and antifungal agents, for example, parabens,chlorobutanol, phenol, sorbic acid, thimerosal, and the like.

Sterile injectable solutions are prepared by incorporating thehigh-purity compositions of Formula 1 in the required amount in theappropriate solvent with various other ingredients as enumerated above,as required, followed by sterile filtration. Generally, dispersions areprepared by incorporating the various sterilized high-puritycompositions of Formula 1 into a sterile vehicle which contains thebasic dispersion medium and the required other ingredients from thoseenumerated above. In the case of sterile powders for the preparation ofsterile injectable solutions, the methods of preparation arevacuum-drying and freeze-drying techniques, which yield a powder of thehigh-purity compositions of Formula 1 plus any additional desiredingredient from a previously sterile-filtered solution thereof.

The high-purity compositions of Formula 1 may be impregnated into astent by diffusion, for example, or coated onto the stent such as in agel form, for example, using procedures known to one of skill in the artin light of the present disclosure.

Oral administration is another route for administration of thehigh-purity compositions of Formula 1. Embodiments include oraladministration via capsule or enteric-coated tablets, or the like, whichprevent degradation of the high-purity compositions of Formula 1 in thestomach. In making the pharmaceutical compositions that include at leasthigh-purity composition of Formula 1, the active ingredient is usuallydiluted by an excipient and/or enclosed within such a carrier that canbe in the form of a capsule, sachet, paper or other container. When theexcipient serves as a diluent, in can be a solid, semi-solid, or liquidmaterial (as above), which acts as a vehicle, carrier or medium for thehigh-purity compositions of Formula 1. Thus, the compositions can be inthe form of tablets, pills, powders, lozenges, sachets, cachets,elixirs, suspensions, emulsions, solutions, syrups, aerosols (as a solidor in a liquid medium), ointments containing, for example, up to 10% byweight of the high-purity compositions of Formula 1, soft and hardgelatin capsules, sterile injectable solutions, and sterile packagedpowders.

Some examples of suitable excipients include lactose, dextrose, sucrose,sorbitol, mannitol, starches, gum acacia, calcium phosphate, alginates,tragacanth, gelatin, calcium silicate, microcrystalline cellulose,polyvinylpyrrolidone, cellulose, sterile water, syrup, andmethylcellulose. The formulations can additionally include: lubricatingagents such as talc, magnesium stearate, and mineral oil; wettingagents; emulsifying and suspending agents; preserving agents such asmethyl- and propylhydroxy-benzoates; sweetening agents; and flavoringagents.

The high-purity compositions of Formula 1 can be formulated so as toprovide quick, sustained or delayed release of the high-puritycompositions of Formula 1 after administration to the patient byemploying procedures known in the art. Controlled release drug deliverysystems for oral administration include osmotic pump systems anddissolutional systems containing polymer-coated reservoirs ordrug-polymer matrix formulations. Examples of controlled release systemsare given in U.S. Pat. Nos. 3,845,770; 4,326,525; 4,902,514; and5,616,345. Another formulation for use in the methods described hereinemploys transdermal delivery devices (“patches”). Such transdermalpatches may be used to provide continuous or discontinuous infusion ofthe high-purity compositions of Formula 1 in controlled amounts, see,e.g., U.S. Pat. Nos. 5,023,252, 4,992,445 and 5,001,139. Such patchesmay be constructed for continuous, pulsatile, or on demand delivery ofpharmaceutical agents.

The compositions are optionally formulated in a unit dosage form. Theterm “unit dosage form(s)” refers to physically discrete units suitableas unitary dosages for human subjects and other mammals, each unitcontaining a predetermined quantity of active material calculated toproduce the desired therapeutic effect, in association with a suitablepharmaceutical excipient (e.g., a tablet, capsule, ampoule). Thehigh-purity compositions of Formula 1 are effective over a wide dosagerange and are generally administered in a pharmaceutically effectiveamount. For oral administration, each dosage unit contains from 10 mg to2 g of a high-purity composition of Formula 1, and for parenteraladministration, from 10 to 700 mg of a high-purity composition ofFormula 1, including about 350 mg. The amount of the compound actuallyadministered will be determined by a physician, in the light of therelevant circumstances, including the condition to be treated, thechosen route of administration, the actual compound administered and itsrelative activity, the age, weight, and response of the individualpatient, the severity of the patient's symptoms, and the like.

For preparing solid compositions such as tablets, the high-puritycomposition of Formula 1 is mixed with a pharmaceutical excipient toform a solid preformulation composition containing a homogeneous mixtureof a high-purity composition of Formula 1. When referring to thesepreformulation compositions as homogeneous, it is meant that the activeingredient is dispersed evenly throughout the composition so that thecomposition may be readily subdivided into equally effective unit dosageforms such as tablets, pills and capsules.

The tablets or pills described herein may be coated or otherwisecompounded to provide a dosage form affording the advantage of prolongedaction, or to protect from the acid conditions of the stomach. Forexample, the tablet or pill can comprise an inner dosage and an outerdosage component, the latter being in the form of an envelope over theformer. The two components can be separated by an enteric layer thatserves to resist disintegration in the stomach and permit the innercomponent to pass intact into the duodenum or to be delayed in release.A variety of materials can be used for such enteric layers or coatings,such materials including a number of polymeric acids and mixtures ofpolymeric acids with such materials as shellac, cetyl alcohol, andcellulose acetate.

Compositions for inhalation or insufflation include solutions andsuspensions in pharmaceutically acceptable, aqueous or organic solvents,or mixtures thereof, and powders. The liquid or solid compositions maycontain suitable pharmaceutically acceptable excipients as describedherein. Such compositions are administered by the oral or nasalrespiratory route for local or systemic effect. Compositions inpharmaceutically acceptable solvents may be nebulized by use of inertgases. Nebulized solutions may be inhaled directly from the nebulizingdevice or the nebulizing device may be attached to a facemask tent, orintermittent positive pressure-breathing machine. Solution, suspension,or powder compositions may be administered, orally or nasally, fromdevices that deliver the formulation in an appropriate manner.

The specific dose will vary depending on the particular high-puritycomposition of Formula 1 chosen, the dosing regimen to be followed, andthe particular therapeutic energy or agent with which it isadministered, employing dosages within the range of about 0.01 mg/kg pertreatment up to about 100 mg/kg per treatment, preferably about 0.1mg/kg per treatment to about 50 mg/kg per treatment. There are specificdifferences in the most effective dosimetry depending on the apicalligands chosen, because of the wide range of properties available, suchas solubilities, lipophilicity properties, lower toxicity, and improvedstability.

Administration for Photodynamic Therapy

By way of example, a high-purity composition of Formula 1, havinglutetium as a metal in the texaphyrin, may be administered in solution,optionally in 5% mannitol USP. Dosages of about 1.0 to 2.0 mg/kg toabout 4.0 to 7.0 mg/kg, including 3.0 mg/kg, are employed, although insome cases a maximum tolerated dose may be higher, for example about 5mg/kg. The texaphyrin is administered by intravenous injection, followedby a waiting period of from as short a time as several minutes or about3 hours to as long as about 72 or 96 hours (depending on the treatmentbeing effected) to facilitate intracellular uptake and clearance fromthe plasma and extracellular matrix prior to the administration ofphotoirradiation.

Dose levels for certain uses may range from about 0.05 mg/kg to about 20mg/kg administered in single or multiple doses (e.g., before eachfraction of photoirradiation). The lower dosage range would beapplicable, for example, to intra-arterial injection or for impregnatedstents.

The optimum length of time between administration of high-puritycompositions of Formula 1 and light treatment can vary depending on themode of administration, the form of administration, and the type oftarget tissue. Typically, the high-purity compositions of Formula 1persists for a period of minutes to hours, depending on the high-puritycomposition of Formula 1, the formulation, the dose, the infusion rate,as well as the type of tissue and tissue size.

When employing photodynamic therapy, a target area is treated withlight, for example at about 740±16.5 nm. After the photosensitizinghigh-purity composition of Formula 1 has been administered, the tissuebeing treated is photo irradiated at a wavelength similar to theabsorbance of the high-purity composition of Formula 1, usually eitherabout 440 to 540 nm or about 700 to 800 nm, or about 450 to 520 nm, orabout 720 to 780 nm, or about 460 to 500 nm or about 725 to 760 nm. Thelight source may be a laser, a light-emitting diode, or filtered lightfrom, for example, a xenon lamp; and the light may be administeredtopically, endoscopically, or interstitially (via, e.g., a fiber opticprobe), or intra-arterially. In one embodiment, the light isadministered using a slit-lamp delivery system. The fluence andirradiance during the photo irradiating treatment can vary depending onthe type of tissue, depth of target tissue, and the amount of overlyingfluid or blood. For example, a total light energy of about 100 J/cm2 canbe delivered at a power of 200 mW to 250 mW, depending upon the targettissue.

Administration with Chemotherapeutic Drugs

High-purity compositions of Formula 1 may be administered before, at thesame time as, or after administration of one or more chemotherapeuticdrugs. The high-purity composition of Formula 1 may be administered as asingle dose, or it may be administered as two or more doses separated byan interval of time. The high-purity composition of Formula 1 may beadministered concurrently with, or from about 1 minute to about 12 hoursfollowing administration of a chemotherapeutic drug, preferably fromabout 5 minutes to about 5 hours, more preferably about 4 to 5 hours.The dosing protocol may be repeated, from one to three times, forexample. A time frame that has been successful in vivo is administrationof a high-purity composition of Formula 1 about 5 minutes and about 5hours after administration of a chemotherapeutic agent, with theprotocol being performed once per week for three weeks. Administrationmay be intra-arterial injection, intravenous, intraperitoneal,intramuscular, subcutaneous, oral, topical, or via a device such as astent.

Administering a high-purity composition of Formula 1 and achemotherapeutic drug to the subject may be prior to, concurrent with,or following vascular intervention. The method may begin at a timeroughly accompanying a vascular intervention, such as an angioplasticprocedure, for example. Multiple or single treatments prior to, at thetime of, or subsequent to the procedure may be used. “Roughlyaccompanying a vascular intervention” refers to a time period within theambit of the effects of the vascular intervention. Typically, an initialdose of a high-purity composition of Formula 1 and chemotherapeutic drugwill be within 6 to 12 hours of the vascular intervention, preferablywithin 6 hours thereafter. Follow-up dosages may be made at weekly,biweekly, or monthly intervals. Design of particular protocols dependson the individual subject, the condition of the subject, the design ofdosage levels, and the judgment of the attending practitioner.

Administration for Radiation Sensitization

High-purity compositions of Formula 1, where the metal is gadolinium,may be administered in a solution containing about 2 mM of the compoundof Formula 1, optionally in 5% mannitol USP/water (sterile andnon-pyrogenic solution); in a further or alternative composition, thesolution contains about 2.2 mM of the compound of Formula 1 (in additionto other components described herein). Dosages of 0.1 mg/kg up to ashigh as about 29.0 mg/kg have been delivered, preferably about 3.0 toabout 15.0 mg/kg (for volume of about 90 to 450 mL) may be employed,optionally with pre-medication using anti-emetics when dosing aboveabout 6.0 mg/kg. The compound is administered via intravenous infusionover about a 5 to 10 minute period, followed by a waiting period ofabout 2 to 5 hours to facilitate intracellular uptake and clearance fromthe plasma and extracellular matrix prior to the administration ofradiation.

When employing whole brain radiation therapy, a course of 30 Gy in ten(10) fractions of radiation may be administered over consecutive daysexcluding weekends and holidays. In the treatment of brain metastases,whole brain megavolt radiation therapy is delivered with 60Coteletherapy or a ≧4 MV linear accelerator with isocenter distances of atleast 80 cm, using isocentric techniques, opposed lateral fields andexclusion of the eyes. A minimum dose rate at the midplane in the brainon the central axis is about 0.5 Gy per minute.

High-purity compositions of Formula 1 used as radiation sensitizers maybe administered before, or at the same time as, or after administrationof the ionizing radiation. The high-purity composition of Formula 1 maybe administered as a single dose, as an infusion, or it may beadministered as two or more doses separated by an interval of time.Where the high-purity composition of Formula 1 is administered as two ormore doses, the time interval between the high-purity composition ofFormula 1 administrations may be from about one minute to a number ofdays, from about 5 minutes to about 1 day, or from about 10 minutes toabout 10 hours. The dosing protocol may be repeated, from one to ten ormore times, for example. Dose levels for radiation sensitization mayrange from about 0.05 mg/kg to about 20 mg/kg administered in single ormultiple doses (e.g. before each fraction of radiation). The lowerdosage range would be preferred for intra-arterial injection or forimpregnated stents.

Administration may be intra-arterial injection, intravenous,intraperitoneal, intramuscular, subcutaneous, oral, topical, or via animpregnated or coated device such as a stent, for example, or anartery-inserted cylindrical polymer. In one aspect of the methods fortreating described herein, a patient having restenosis or at risk forrestenosis is administered a dose of high-purity composition of Formula1 at intervals with each dose of photoirradiation.

Administering a high-purity composition of Formula 1 to the subject maybe prior to, concurrent with, or following vascular intervention, andthe intervention is followed by photoirradiation. The method may beginprior to, such as about 24 to 48 hours prior to, or at a time roughlyaccompanying vascular intervention, for example. Multiple or singletreatments prior to, at the time of, or subsequent to the procedure maybe used. “Roughly accompanying the vascular intervention” refers to atime period within the ambit of the effects of the vascularintervention. Typically, an initial dose of a high-purity composition ofFormula 1 and light will be within 1 to 24 hours of the vascularintervention, preferably within about 5 to 24 hours thereafter.Follow-up dosages may be made at weekly, biweekly, monthly or longerintervals. Design of particular protocols depends on the individualsubject, the condition of the subject, the design of dosage levels, andthe judgment of the attending practitioner.

Administration for Sonodynamic Therapy

The use of texaphyrins in sonodynamic therapy is described in U.S.patent application Ser. No. 09/111,148, which was converted to U.S.Provisional Application Ser. No. 60/155,256, from which a continuationwas filed on Jan. 5, 2001, having U.S. patent application Ser. No.09/755,824, now abandoned, which is incorporated herein by reference.Texaphyrin is administered before administration of the ultrasound. Thetexaphyrin may be administered as a single dose, or it may beadministered as two or more doses separated by an interval of time.Parenteral administration is typical, including by intravenous andinterarterial injection. Other common routes of administration can alsobe employed.

Ultrasound is generated by a focused array transducer driven by a poweramplifier. The transducer can vary in diameter and spherical curvatureto allow for variation of the focus of the ultrasonic output.Commercially available therapeutic ultrasound devices may be employed inthe practice of such a method. The duration and wave frequency,including the type of wave employed may vary, and the preferred durationof treatment will vary from case to case within the judgment of thetreating physician. Both progressive wave mode patterns and standingwave patterns have been successful in producing cavitation of diseasedtissue. When using progressive waves, the second harmonic canadvantageously be superimposed onto the fundamental wave. Types ofultrasound employed in such a method are ultrasound of low intensity,non-thermal ultrasound, i.e., ultrasound generated within thewavelengths of about 0.1 MHz and 5.0 MHz and at intensities betweenabout 3.0 and 5.0 W/cm2.

In further applications, the compound of Formula 1, where M is Gd+3, maybe administered with NAD(P)H, ascorbate and other reducing agents underapproximate physiological conditions, leading to reactive oxygen speciesgeneration. Depletion of these reducing agents will inhibit biochemicalpathways that in vivo utilize reducing agents to effect repair of thedamage inflicted by reactive oxygen species. Such a method may be usedto treat cancer and cardiovascular diseases. See U.S. Pat. No.6,825,186, which is incorporated by reference in its entirety. Theability of compositions comprising a compound of Formula 1 to catalyzethe oxidation of these reducing agents is expected to improve as thepurity of the composition improves; hence, the high-purity compositionsdescribed herein are expected to result in an improved catalytic, andtherapeutic, effectiveness.

While embodiments have been shown and described herein, it will beobvious to those skilled in the art that such embodiments are providedby way of example only. Numerous variations, changes, and substitutionswill now occur to those skilled in the art without departing from theinvention. It should be understood that various alternatives to theembodiments described herein may be employed in practicing theinvention. It is intended that the following claims define the scope ofthe invention and that methods and structures within the scope of theseclaims and their equivalents be covered thereby.

EXAMPLES

All solvents and reagents were of reagent grade quality, purchasedcommercially, and used as received.

Example 1 Synthesis of a Compound of Formula 7 x=3

A solution of compound of Formula 8 (x=3; 74 g) in tetrahydrofuran (39g) was added to an aqueous solution of sodium hydroxide (25%, 235 g),maintaining the temperature at 0-10° C. (see FIG. 1). It is important tomaintain the temperature within this range. A solution ofp-toluenesulfonyl chloride (TsCl, 78 g) in tetrahydrofuran (126 g) wasadded to compound of Formula 8 over 4 hours, also at 0-10° C. Thereaction mixture was stirred for up to 90 minutes and neutralized by anaddition of dilute hydrochloric acid (33%, 115 g) at 0-10° C. The loweraqueous layer was discarded and the organic layer was washed twice withbrine (24%, 55 ml). The resulting solution of compound of Formula 7 intetrahydrofuran is used directly in the next step.

Example 2 Synthesis of a Compound of Formula 5 x=3; R1=R2=H

o-Dihydroxybenzene (25 g), potassium carbonate (61 g) and the solutionof compound of Formula 7 (x=3) were mixed and heated at reflux until≦0.5% compound of Formula 7 remained (24-28 hr) (see FIG. 1). Thereaction mixture was filtered and the filter cake was washed twice withtetrahydrofuran (92 g). The combined filtrates were concentrated underreduced pressure and a temperature of up to 110° C. The residue wascooled and treated with aqueous sodium hydroxide solution (6.5%, 315 g)at 20-30° C. Dichloromethane (106 ml) was added and the mixture agitatedand separated. The organic phase was washed with water (150 ml) anddistilled to dryness at up to 100° C. at atmospheric pressure. Theresidue was dissolved in acetic acid (˜60 g) and used as an acetic acidsolution in the next step.

Example 3 Synthesis of a Compound of Formula 4 x=3; R1=R2=H

A solution of compound of Formula 5 (x=3; R1=R2=H) in acetic acid wasadded to 90% nitric acid (580 g) at a temperature of 0-5° C. Thereaction was stirred for 30-60 minutes at this temperature, thenquenched by addition to water (620 ml) at 0-10° C. The reaction mixturewas extracted twice with dichloromethane (265 ml followed by 130 ml),and the combined organic phases was washed with aqueous sodium hydroxidesolution (5%, about 360 kg) at pH 5.5-8.5 and then with water (350 ml).The dichloromethane was distilled under reduced pressure and wasreplaced by IPA (50 g), which was also distilled, leaving the crudecompound of Formula 4 as oil at 50-60° C. The crude compound of Formula4 was added slowly to IPA (600 g) at 2-6° C. (critical parameter) andthe mixture was held at this temperature for at least 1 hr. Theprecipitated compound of Formula 4 was filtered and washed with coldIPA. The wet cake was melted at 50-60° C. and the crystallizationprocedure was repeated. The wet cake was then dried at 25° C. underreduced pressure. ¹H NMR (CDCl₃): δ 3.37 (6H, s, OCH3), 3.53-3.56 (4H,m, PEG group), 3.62-3.64 (4H, m, PEG group), 3.65-3.67 (4H, m, PEGgroup), 3.71-3.74 (4H, m, PEG group), 3.92 (4H, t, PEG group), 4.31 (4H,t, PEG group), 7.49 (2H, s, aromatic). ¹³C NMR (CDCl₃): δ 58.93, 69.53,70.01, 70.60, 70.69, 71.05, 71.94, 109.56, 136.73, 151.81. LC/MS (APCImode), MH+: m/e 493.2. Elemental analysis: calcd. for C20H32N2O12: C,48.78; H, 6.55; N, 5.69. Found: C, 48.77; H, 6.55; N, 5.54.

What is claimed is:
 1. A high-purity composition comprising a compoundaccording to Formula 1

wherein: M is a trivalent metal cation selected from the groupconsisting of Gd⁺³, and Lu⁺³; each X is independently selected from thegroup consisting of OH⁻, AcO⁻, Cl⁻, Br⁻, I⁻, F⁻, H₂PO₄ ⁻, ClO⁻, ClO₂ ⁻,ClO₃ ⁻, ClO₄ ⁻, HCO₃ ⁻, HSO₄ ⁻, NO₃ ⁻, N₃ ⁻, CN⁻, SCN⁻, and OCN⁻ R₃, R₄,R₅, R₆, R₇ and R₈ are independently H, OH, C_(n)H_((2n+1))O_(y) orOC_(n)H_((2n+1))O_(y) and R₁, R₂ are independently H or C₁-C₆ alkylwhere at least one of R₃, R₄, R₅, R₆, R₇ and R₈ is C_(n)H_((2n+1))O_(y)or OC_(n)H_((2n+1))O_(y), having at least one hydroxyl substituent; n isa positive integer from 1 to 11; y is zero or a positive integer lessthan or equal to n; each x is independently selected from the groupconsisting of 2, 3, 4, 5, and 6; wherein at least about 98.4% ofcompounds of Formula 1 in the composition have the same structure. 2.The composition of claim 1 wherein M is Gd⁺³.
 3. The composition ofclaim 2 wherein R₄ and R₇ are C₃H₆OH; R₅ and R₆ are C₂H₅; R₃ and R₈ areCH₃; R₁ and R₂ are H.
 4. The composition of claim 3 wherein each x is 3.5. The composition of claim 4 wherein each X is AcO⁻.
 6. The compositionof claim 1 wherein at least about 99.3% of the compounds of Formula 1 inthe purified composition have the same molecular weight.
 7. Thecomposition of claim 1 wherein at least about 99.3% of the compounds ofFormula 1 in the purified composition have the same structure.
 8. Amethod for synthesizing a purified sample of a compound of Formula 4

wherein R₁ and R₂ are independently H or C₁-C₆ alkyl; each x isindependently selected from the group consisting of 2, 3, 4, 5, and 6;and wherein at least about 99% of the compounds of Formula 4 in thepurified sample have the same structure comprising: reacting a compoundof Formula 5

with about 90% HNO₃ in AcOH at about 0-5° C. for at least 30 minutes,wherein at least about 99% of the compound of Formula 4 formed in thepurified sample has the same molecular weight.
 9. The method of claim 8wherein each x is
 3. 10. The method of claim 8 wherein at least about99.3% of the compounds of Formula 4 in the purified sample have the samestructure.
 11. The method of claim 8, wherein the compound of Formula 5is provided as a purified sample synthesized by a method comprising:reacting tosylated compound of Formula 7

with o-dihydroxybenzene of Formula 6

in the presence of K₂CO₃ and an aprotic solvent and heating the mixtureat about 65° C. for about 24 hr, wherein at least about 98.4% of thecompounds of Formula 5 formed in the purified sample have the samestructure.
 12. The method of claim 11 wherein said aprotic solvent isselected from the group consisting of aldehydes, ketones, dimethylsulfoxide, dimethyl formamide, pyridine, diethyl ether, ethyl acetate,propyl acetate, isopropyl acetate, butyl acetate, acetonitrile,benzonitrile, sulfur dioxide, tetrahydrofuran, and hexamethylphosphoramide.
 13. The method of claim 12 wherein said aprotic solventis tetrahydrofuran.
 14. The method of claim 11 wherein each x is
 3. 15.The method of claim 11 wherein at least about 99.3% of the compounds ofFormula 5 in the purified sample have the same structure.
 16. Apharmaceutical composition comprising a compound according to Formula 1

wherein: M is a trivalent metal cation selected from the groupconsisting of Gd⁺³, and Lu⁺³; each X is independently selected from thegroup consisting of OH⁻, AcO⁻, Cl⁻, Br⁻, I⁻, F⁻, H₂PO₄ ⁻, ClO⁻, ClO₂ ⁻,ClO₃ ⁻, ClO₄ ⁻, HCO₃ ⁻, HSO₄ ⁻, NO₃ ⁻, N₃ ⁻, CN⁻, SCN⁻, and OCN⁻; R₃,R₄, R₅, R₆, R₇ and R₈ are independently H, OH, C_(n)H_((2n+1))O_(y) orOC_(n)H_((2n+1))O_(y) and R₁, R₂ are independently H or C₁-C₆ alkylwhere at least one of R₃, R₄, R₅, R₆, R₇ and R₈ is C_(n)H_((2n+1))O_(y)or OC_(n)H_((2n+1))O_(y), having at least one hydroxy substituent; n isa positive integer from 1 to 11; y is zero or a positive integer lessthan or equal to n; each x is independently selected from the groupconsisting of 2, 3, 4, 5, and 6; and a pharmaceutically acceptablecarrier, wherein at least about 98.4% of the compounds of Formula 1 inthe pharmaceutical composition have the same structure.
 17. Thepharmaceutical composition of claim 16 wherein M is Gd⁺³.
 18. Thepharmaceutical composition of claim 17 wherein R₄ and R₇ are C₃H₆OH; R₅and R₆ are C₂H₅; R₃ and R₈ are CH₃; R₁ and R₂ are H.
 19. Thepharmaceutical composition of claim 18 wherein each x is
 3. 20. Thepharmaceutical composition of claim 19 wherein each X is AcO⁻.
 21. Thepharmaceutical composition of claim 16 wherein at least about 99.3% ofthe compounds of Formula 1 in the pharmaceutical composition have thesame molecular weight.
 22. The pharmaceutical composition of claim 16wherein at least about 99.3% of the compound of Formula 1 in thepharmaceutical composition have the same structure.
 23. Thepharmaceutical composition of claim 16 wherein the pharmaceuticallyacceptable carrier is suitable for parenteral administration.
 24. Amethod of treating a cancer comprising administering to a human patientin need thereof an effective amount of a purified composition comprisinga compound of Formula 1

wherein: M is a trivalent metal cation selected from the groupconsisting of Gd⁺³, and Lu⁺³; X is independently selected from the groupconsisting of OH⁻, AcO⁻, Cl⁻, Br⁻, I⁻, F⁻, H₂PO₄ ⁻, ClO⁻, ClO₂ ⁻, ClO₃⁻, ClO₄ ⁻, HCO₃ ⁻, HSO₄ ⁻, NO₃ ⁻, N₃ ⁻, CN⁻, SCN⁻, and OCN⁻; R₃, R₄, R₅,R₆, R₇ and R₈ are independently H, OH, C_(n)H_((2n+1))O_(y) orOC_(n)H_((2n+1))O_(y) and R₁, R₂ are independently H or C₁-C₆ alkylwhere at least one of R₃, R₄, R₅, R₆, R₇ and R₈ is C_(n)H_((2n+1))O_(y)or OC_(n)H_((2n+1))O_(y), having at least one hydroxyl substituent; n isa positive integer from 1 to 11; y is zero or a positive integer lessthan or equal to n; each x is independently selected from the groupconsisting of 2, 3, 4, 5, and 6; and wherein at least about 98.4% of thecompounds of Formula 1 in the purified composition have the samemolecular weight; or a pharmaceutically acceptable salt, solvate,isomers, tautomers, metabolites, analogs, or prodrugs thereof.
 25. Themethod of claim 24 wherein M is Gd⁺³.
 26. The method of claim 25 whereinR₄ and R₇ are C₃H₆OH; R₅ and R₆ are C₂H₅; R₃ and R₈ are CH₃; R₁ and R₂are H.
 27. The method of claim 26 wherein each x is
 3. 28. The method ofclaim 27 wherein each X is AcO⁻.
 29. The method of claim 24 wherein atleast about 99.3% of the compounds of Formula 1 in the purified samplehave the same molecular weight.
 30. The method of claim 24 wherein atleast about 99.3% of the compounds of Formula 1 in the purified samplehave the same structure.
 31. The method of claim 24 further comprisingproviding to the human patient an additional therapy selected from thegroup consisting of surgery, radiation therapy, chemotherapy, genetherapy, immunotherapy, or a combination thereof.
 32. The method ofclaim 24 wherein the cancer is metastatic brain cancer.